Solea Senegalensis Juveniles Research Articles

Effects of acute exposure to exogenous ammonia on cerebral monoaminergic neurotransmitters in juvenile Solea senegalensis

The present study explored the potential role of brain catecholaminergic and serotoninergic systems as neuronal targets for the toxicological effects of acute ammonia exposure (0.28 mg l(-1) of un-ionized ammonia for 12 and 24 h) in juvenile sole (Solea senegalensis). In addition, plasma cortisol levels were measured. The results showed significant increases in their concentrations that were similar after 12 and 24 h of exposure. These data indicate that acute exposure (12 and 24 h) to ammonia initiates a typical stress response in the Senegalese sole, with stimulation of the hypothalamus-pituitary-interrenal axis. The concentrations of dopamine (DA), serotonin (5-hydroxytryptamine; 5-HT) and noradrenaline (NA), and their metabolites, 3, 4-dihydroxyphenylacetic acid (DOPAC) and 5-hydroxy-3-indoleacetic acid (5HIAA), were measured in the hypothalamus, telencephalon and optic tectum. The main changes induced by acute exposure to ammonia were decreases in the concentrations of 5-HT and DA, which were significant in most of the brain regions studied. The ratios of 5-HIAA/5-HT and DOPAC/DA increased in all regions and at all times studied, although in the case of the DOPAC/DA ratio, the increases were only significant in the hypothalamus (24 h exposure) and in the optic tectum (12 and 24 h exposure). These changes indicated that exposure to ammonia elicited time-dependent increases in serotoninergic and dopaminergic activity in the hypothalamus, telencephalon and optic tectum.

Effects of acute exposure to 2-phenoxyethanol, clove oil, MS-222, and metomidate on primary and secondary stress responses in Senegalese sole ( Solea senegalensis Kaup 1858)

The present study investigated possible stress effects in juvenile Solea senegalensis caused by exposure for 5, 10, 20 and 30 min to the anaesthetic agents 2-phenoxyethanol (500 mg L − 1 ), clove oil (80 mg L − 1 ), MS-222 (200 mg L − 1 ) and metomidate (6 mg L − 1 ). The effects were also assessed 24 h after the longest exposure (30 min), and after killing the fish with a sharp blow to the head. Levels of cortisol, haematocrit, haemoglobin, glucose and lactate were measured in the fish as indicators of stress. The plasma concentrations of cortisol increased significantly after 20 min exposure to 2-phenoxyethanol, and after 20 and 30 min exposure to clove oil and MS-222. The greatest increases occurred after 30 min exposure to clove oil and MS-222, and 20 min exposure to 2-phenoxyethanol. With metomidate, significant increases in cortisol levels were only observed after 10 min exposure, and the magnitude of the increases was much lower than after exposure to the other anaesthetics. None of the anaesthetic agents induced significant changes in the haematocrit or haemoglobin levels. By contrast, the concentration of glucose increased significantly after 10, 20 and 30 min exposure to MS-222, after 20 and 30 min exposure to clove oil and after 30 min exposure to 2-phenoxyethanol and to metomidate. The concentrations of lactate increased significantly after 20 and 30 min exposure to clove oil and to MS-222, and after 30 min exposure to 2-phenoxyethanol and to metomidate. In all cases the levels of the stress indicators returned to control values (5 min exposure), 24 h after 30 min exposure to the anaesthetics. The results indicate that exposure to the anaesthetics induces a stress effect, characterised by stimulation of the hypothalamus–pituitary–interrenal (HPI) axis and to a much lesser extent, the sympathetic nervous system-associated chromaffin tissue.

Proliferating cells in suborbital tissue drive eye migration in flatfish

The left/right asymmetry of adult flatfishes (Pleuronectiformes) is remarkable given the external body symmetry of the larval fish. The best-known change is the migration of their eyes: one eye migrates from one side to the other. Two extinct primitive pleuronectiformes with incomplete orbital migration have again attracted public attention to the mechanism of eye migration, a subject of speculation and research for over a century. Cranial asymmetry is currently believed to be responsible for eye migration. Contrary to that hypothesis, we show here that the initial migration of the eye is caused by cell proliferation in the suborbital tissue of the blind side and that the twist of frontal bone is dependent on eye migration. The inhibition of cell proliferation in the suborbital area of the blind side by microinjected colchicine was able to prevent eye migration and, thereafter, cranial asymmetry in juvenile Solea senegalensis (right sideness, Soleidae), Cynoglossus semilaevis (left sideness, Cynoglossidae), and Paralichthys olivaceus (left sideness, Paralichthyidae) with a bottom-dwelling lifestyle. Our results correct the current misunderstanding that eye migration is driven by the cranial asymmetry and simplify the explanation for broken left/right eye-symmetry. Our findings should help to focus the search on eye migration-related genes associated with cell proliferation. Finally, a novel model is proposed in this research which provides a reasonable explanation for differences in the migrating eye between, and sometimes within, different species of flatfish and which should aid in our overall understanding of eye migration in the ontogenesis and evolution of Pleuronectiformes.

A description of chloride cell and kidney tubule alterations in the flatfish Solea senegalensis exposed to moderately contaminated sediments from the Sado estuary (Portugal)

The effects of sediment-bound contaminants on kidney and gill chloride cells were surveyed in juvenile Solea senegalensis exposed to fresh sediments collected from three distinct sites of the Sado Estuary (Portugal) in a 28-day laboratorial assay. Sediments were analyzed for metallic contaminants, polycyclic aromatic hydrocarbons and organochlorines as well as for total organic matter, redox potential and fine fraction. The potential for causing adverse biological effects of each surveyed sediment was assessed by comparison of contaminant levels to available guidelines for coastal sediments, namely the Threshold Effects Level ( TEL) and the Probable Effects Level ( PEL). The Sediment Quality Guideline Quotient indices ( SQGQ) were calculated to compare the overall contamination levels of the three stations. A qualitative approach was employed to analyze the histo/cytopathological traits in gill chloride cells and body kidney of fish exposed to each tested sediment for 0, 14 and 28 days. The results showed that sediment contamination can be considered low to moderate and that the least contaminated sediment (from a reference site, with the lowest SQGQ) caused lesser changes in the surveyed organs. However, the most contaminated sediment (by both metallic and organic xenobiotics, with highest SQGQ) was neither responsible for the highest mortality nor for the most pronounced lesions. Exposure to the sediment presenting an intermediate SQGQ, essentially contaminated by organic compounds, caused the highest mortality (48%) and the most severe damage to kidneys, up to full renal necrosis. Chloride cell alterations were similar in fish exposed to the two most contaminated sediments and consisted of a pronounced cellular hypertrophy, likely involving fluid retention and loss of mitochondria. It can be concluded that sediment contamination considered to be low or moderate may be responsible for severe injury to cells and parenchyma involved in the maintenance of osmotic balance, contributing for the high mortality levels observed. The results suggest that sediment-bound organic contaminants such as PAHs (polycyclic aromatic hydrocarbons) and PCBs (polychlorinated biphenyls) may be very toxic to the analyzed organs, especially the kidney, even when present in low-risk concentrations.

Lack of essential fatty acids in live feed during larval and post-larval rearing: effect on the performance of juvenile Solea senegalensis

Despite the large progress obtained in recent years, Senegalese sole (Solea senegalensis) production of high quality juveniles is still a bottleneck. This paper examines the effect of larval and post-larval lipid nutrition on juvenile performance and quality. Four dietary treatments were tested: A—enriched Artemia spp. (EA); B—non-enriched Artemia spp. (NEA); C—EA during the pelagic larval period and NEA after larval settlement; D—50% EA and 50% NEA. Juvenile fatty acid profile at 60 days after hatching (DAH) clearly reflected the larval and post-larval diet composition. Feeding sole larvae on NEA (poor in lipids and essential fatty acids-EFA) had a negative effect, reducing growth (total length and dry weight) after 30 DAH and decreasing digestive enzyme activity at the end of the rearing period (60 DAH). However, relatively good performance compared to the EFA-richest treatment (A) was obtained when larvae were fed 50% EA and 50% NEA (D) or even EA only during the pelagic larval period followed by NEA after larval settlement (C). Malpigmentation was not affected by the dietary regimes and its incidence was very low. However, skeletal deformities were prevalent, particularly in the caudal complex, independently of diet. The results confirm that Senegalese sole appear to have lower larval EFA requirements than most cultured marine species and potentially even lower requirements during the post-larval stage. The importance of studying the impact of early nutrition on later juvenile stages was clearly highlighted in this study.

Biochemical endpoints on juvenile Solea senegalensis exposed to estuarine sediments: the effect of contaminant mixtures on metallothionein and CYP1A induction

Juvenile Solea senegalensis were exposed to fresh sediments from three stations of the Sado estuary (Portugal) in 28-day laboratory assays. Sediments revealed distinct levels of total organic matter, fine fraction, redox potential, trace elements (arsenic, cadmium, chromium, copper, nickel, lead and zinc) and organic contaminants (polycyclic aromatic hydrocarbons, polychlorinated biphenyls and a pesticide: dichloro diphenyl trichloroethane). Organisms were surveyed for contaminant bioaccumulation and induction of two hepatic biochemical biomarkers: metallothionein (MT) and cytochrome P450 (CYP1A), as potential indicators of exposure to metallic and organic contaminants, respectively. Using an integrative approach it was established that, although bioaccumulation is in general accordance with sediment contamination, lethality and biomarker responses are not linearly dependent of the cumulative concentrations of sediment contaminants but rather of their bioavailability and synergistic effects in organisms. It is concluded that metals and organic contaminants modulate both MT and CYP1A induction and it is suggested that reactive oxygen species may be the link between responses and effects of toxicity.

Histological biomarkers in liver and gills of juvenile Solea senegalensis exposed to contaminated estuarine sediments: A weighted indices approach

Young juvenile Solea senegalensis were exposed to three sediments with distinct contamination profiles collected from a Portuguese estuary subjected to anthropogenic sources of contamination (the Sado estuary, western Portugal). Sediments were surveyed for metals (cadmium, chromium, copper, nickel, lead and zinc), a metalloid (arsenic) and organic contaminants (polycyclic aromatic hydrocarbons, polychlorinated biphenyls and a pesticide, dichloro-diphenyl-trichloroethane plus its metabolites), as well as total organic matter, redox potential and particle fine fraction. The fish were exposed to freshly collected sediments in a 28-day laboratorial assay and collected for histological analyses at days 0 (T 0), 14 (T 14) and 28 (T 28). Individual weighted histopathological indices were obtained, based on presence/absence data of eight and nine liver and gill pathologies, respectively, and on their biological significance. Although livers sustained more severe lesions, the sediments essentially contaminated by organic substances caused more damage to both organs than the sediments contaminated by both metallic and organic contaminants, suggesting a possible synergistic effect. Correlation analyses showed that some alterations are linked, forming distinctive histopathological patterns that are in accordance with the severity of lesions and sediment characteristics. The presence of large eosinophilic bodies in liver and degeneration of mucous cells in gills (a first-time described alteration) were some of the most noticeable alterations observed and were related to sediment organic contaminants. Body size has been found to be negatively correlated with histopathological damage in livers following longer term exposures. It is concluded that histopathological indices provide reliable and discriminatory data even when biomonitoring as complex media as natural sediments. It is also concluded that the effects of contamination may result not only from toxicant concentrations but also from their interactions, relative potency and sediment characteristics that ultimately determine bioavailability.

Effect of copper exposure on growth, condition indices and biomarker response in juvenile sole <i>Solea senegalensis</i>

Juvenile Solea senegalensis were exposed to different concentrations of waterborne copper (Cu) for 15 days in static conditions with continuous aerated artificial salt water at 20°C (± 0.8°C) with a normal photoperiod (10 h/14 h light/dark) and daily feeding. Several measures of exposure and effects were determined: 1) biomarkers - metallothioneins and lipid peroxidation level; 2) mass indices - growth rate and morphometric condition indices; and 3) biochemical condition indices - RNA:DNA ratio and lipid and protein content in fish tissues. Copper exposure triggered the response of the biomarkers and resulted in reduced growth and condition (RNA:DNA and lipid content), but the morphometric indices did not vary. The physiological costs of Cu contamination on condition suggested that lipid reserves were allocated as an energy source to enable exposed fish to respond to Cu toxicity as well as to maintain positive growth rates and protein synthesis throughout the experiment, although with lower growth rates than the control fish. This study showed the importance of selecting suitable biomarkers according to contaminant source, fish species and their life-history stage. In addition, the use of several biomarkers of exposure, growth and specific condition indices can improve fish health determination and should be considered in evaluations of the effects of environmental contaminants on fish.

Small-scale distribution of Solea solea and Solea senegalensis juveniles in the Tagus estuary (Portugal)

The distribution of Solea solea and Solea senegalensis in the Tagus estuary was studied following a small-scale approach. Preliminary sampling revealed that sole concentrated in two areas within their nursery grounds, the main subtidal channel and a large intertidal mudflat. Beam trawls were conducted intensively in the two areas in July 2006. Depth, salinity and water temperature were measured. Substrate samples were collected for sediment type determination and macrobenthos identification and quantification. Generalized linear models were applied in order to explain the occurrence and variability of soles' densities, using depth, salinity, water temperature and abundance of polychaetes, oligochaetes, amphipods, isopods and bivalves as explanatory variables. While S. solea was more abundant in the main subtidal channel, a deeper, warmer and lower salinity area, S. senegalensis abundance was highest at the intertidal mudflat area. Presence of both species in the two areas was associated with abundance of polychaetes (generally with another variable associated), and for S. senegalensis in the subtidal channel it was associated with amphipods and depth. Abundance of S. solea in the main subtidal channel was associated mainly with polychaetes abundance, while that of S. senegalensis was associated with amphipods density. In the intertidal mudflat, bivalves and polychaetes presented significant relationships with both species densities. Some of the factors that had been reported to be important for the distribution of these species in previous studies also do so at a finer scale; however, this small-scale approach provided an in-depth knowledge on habitat selection and spatial segregation of these species within this nursery area.

An early approach for the evaluation of repair processes in fish after exposure to sediment contaminated by an oil spill

A chronic bioassay was carried out under laboratory conditions using juvenile Solea senegalensis to determine the toxicity of contaminants from an oil spill(Prestige). Also, the repair processes in fish affected by contaminants due to oil exposure were evaluated. Over 30 days individuals were exposed to clean sediment (control) and to sediment contaminated by a mixture of polyaromatic hydrocarbons (PAHs) and other substances. The physicochemical parameters of the tanks (salinity, temperature, pH and dissolved oxygen) were controlled during the exposure period. Clean sediment from the Bay of Cádiz (Spain) was used as negative control and was mixed with fuel oil to prepare the dilution (0.5% w:w dry-weight). After the exposure period, fish were labeled and transferred to “clean tanks” (tanks without sediment) in order to study the recovery and the repair processes in the exposed organisms. A biomarker of exposure (ethoxyresorufin-O-deethylase activity - EROD activity) and a biomarker of effect (histopathology) were analyzed during the exposure and recovery period. After 10, 20 and 30 days of exposure, individuals showed significant induction (P < 0.05) of the EROD activity and also presented diverse histopathological damages. The analysis of both the biomarkers of exposure and effect, after the 5th and 10th day of recovery in the “clean tank”, enabled a first evaluation of the repair process of the induced damages due to the fuel oil exposure. After the recovery phase, control individuals showed a more significant decrease (P < 0.05) of the alteration of the measured biomarkers than in the oil-exposed fish. While in the oil-exposed fish the EROD activity showed some recovery, the histopathological damages did hardly improve. According to our results, tissue repair processes probably need longer recovery periods to observe significant improvement of the affected organs. This will be further investigated in the future.

Genotoxic damage in Solea senegalensis exposed to sediments from the Sado Estuary (Portugal): Effects of metallic and organic contaminants

Juvenile Solea senegalensis (Senegalese sole) were exposed to freshly collected sediments from three sites of the Sado Estuary (West-Portuguese coast) in 28-day laboratory assays in order to assess the ecological risk from sediment contaminants, by measuring two genotoxicity biomarkers in peripheral blood: the percentage of Erythrocyte Nuclear Abnormalities (ENA) by use of an adaptation of the micronucleus test, and the percentage of DNA strand-breakage (DNA-SB) with the Comet assay. Sediments were surveyed for metallic (Cr, Ni, Cu, Zn, As, Cd and Pb) and organic (PAHs (polycyclic aromatic hydrocarbons), PCBs (polychlorinated biphenyls) and DDTs (dichloro-diphenyl-trichloroethane)) contaminants. Sediments from site A (farthest from hotspots of contamination) were found to be the least contaminated and weaker inducers of genotoxic damage, whereas sediments from sites B (urban influence) and C (affected by industrial effluents and agricultural runoffs) were responsible for a very significant increase in both ENA and DNA-SB, site B being most contaminated with metals and site C mainly with organic pollutants, especially PAHs and PCBs . Analysis of genotoxic effects showed a strong correlation between the concentrations of PAHs and PCBs and both biomarkers at sampling times T 14 and T 28, while the amounts of Cu, As, Cd and Pb were less strongly correlated, and at T 28 only, with ENA and DNA-SB. These results show that organic contaminants in sediment are stronger and faster acting genotoxic stressors. The results also suggest that metals may have an inhibitory effect on genotoxicity when interacting with organic contaminants, at least during early exposure. ENA and DNA-SB do not show a linear relationship, but a strong correlation exists between the overall increase in genotoxicity caused by exposure to sediment, confirming that they are different, and possibly non-linked effects that respond similarly to exposure. Although the Comet assay showed enhanced sensitivity, the two analyses are complementary and suitable for the biomonitoring of sediment contaminants in a benthic species like S. senegalensis.

Nursery fidelity, food web interactions and primary sources of nutrition of the juveniles of Solea solea and S. senegalensis in the Tagus estuary (Portugal): A stable isotope approach

Stable carbon and nitrogen isotopes were used to assess site fidelity of Solea solea and Solea senegalensis juveniles, to investigate food web interactions and to determine the dominant nutrient pathways in two nursery areas in the Tagus estuary, Portugal. Samples of water from the main sources and from the nursery areas and respective saltmarsh creeks were collected for isotope analysis, as well as sediment, benthic microalgae, saltmarsh halophytes, S. solea, S. senegalensis and its main prey, Nereis diversicolor, Scrobicularia plana and Corophium spp. While site fidelity was high in 0-group juveniles, it was lower for 1-group juveniles, possibly due to an increase in mobility and energy demands with increasing size. Analysis of the food web revealed a complex net of relations. Particulate organic matter from the freshwater sources, from each nursery's waters and saltmarsh creeks presented similar isotopic composition. Sediment isotopic composition and saltmarsh halophytes also did not differentiate the two areas. All components of the food web from the benthic microalgae upwards were isotopically different between the nursery areas. These components were always more enriched in δ 13C and δ 15N at the lower nursery area than at the nursery located upstream, appearing as if there were two parallel trophic chains with little trophic interaction between each other. A mixture of carbon and nitrogen sources is probably being incorporated into the food web. The lower nursery area is more dependent upon an isotopically enriched energy pathway, composed of marine particulate organic matter, marine benthic microalgae and detritus of the C 4 saltmarsh halophyte Spartina maritima. The two nursery areas present a different level of dependence upon the freshwater and marine energy pathways, due to hydrological features, which should be taken into account for S. solea and S. senegalensis fisheries and habitat management.

4581739 Electronically selectable redundant array (ESRA)

Juvenile Solea senegalensis (Senegalese sole) were exposed to freshly collected sediments from three sites of the Sado Estuary (West-Portuguese coast) in 28-day laboratory assays in order to assess the ecological risk from sediment contaminants, by measuring two genotoxicity biomarkers in peripheral blood: the percentage of Erythrocyte Nuclear Abnormalities (ENA) by use of an adaptation of the micronucleus test, and the percentage of DNA strand-breakage (DNA-SB) with the Comet assay. Sediments were surveyed for metallic (Cr, Ni, Cu, Zn, As, Cd and Pb) and organic (PAHs (polycyclic aromatic hydrocarbons), PCBs (polychlorinated biphenyls) and DDTs (dichloro-diphenyl-trichloroethane)) contaminants. Sediments from site A (farthest from hotspots of contamination) were found to be the least contaminated and weaker inducers of genotoxic damage, whereas sediments from sites B (urban influence) and C (affected by industrial effluents and agricultural runoffs) were responsible for a very significant increase in both ENA and DNA-SB, site B being most contaminated with metals and site C mainly with organic pollutants, especially PAHs and PCBs . Analysis of genotoxic effects showed a strong correlation between the concentrations of PAHs and PCBs and both biomarkers at sampling times T14 and T28, while the amounts of Cu, As, Cd and Pb were less strongly correlated, and at T28 only, with ENA and DNA-SB. These results show that organic contaminants in sediment are stronger and faster acting genotoxic stressors. The results also suggest that metals may have an inhibitory effect on genotoxicity when interacting with organic contaminants, at least during early exposure. ENA and DNA-SB do not show a linear relationship, but a strong correlation exists between the overall increase in genotoxicity caused by exposure to sediment, confirming that they are different, and possibly non-linked effects that respond similarly to exposure. Although the Comet assay showed enhanced sensitivity, the two analyses are complementary and suitable for the biomonitoring of sediment contaminants in a benthic species like S. senegalensis.